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Analyzing ATM Function by Electroporation of Endonucleases and Immunofluorescence Microscopy

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ATM Kinase

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1599))

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Abstract

Ataxia-telangiectasia mutated (ATM) protein, which plays a crucial role in DNA damage checkpoint signaling, is activated by DNA double strand breaks (DSBs) caused by ionizing radiation. While radiation exposure induces various types of DNA break ends, here, we describe a method, which enables creating defined types of DSBs by applying restriction endonucleases and foci analysis by immunofluorescence microscopy. The protocol greatly improves our knowledge on specific roles of ATM function in different DNA repair pathways.

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Acknowledgments

This study was supported in part by the Global Center Of Excellence (GCOE) Program and Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Authors and Affiliations

  1. Atomic Bomb Disease Institute, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan

    Keiji Suzuki

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  1. Keiji Suzuki
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Corresponding author

Correspondence to Keiji Suzuki .

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Editors and Affiliations

  1. University of Queensland Centre for Clinical Research (UQCCR), University of Queensland, Herston, Queensland, Australia

    Sergei V. Kozlov

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Suzuki, K. (2017). Analyzing ATM Function by Electroporation of Endonucleases and Immunofluorescence Microscopy. In: Kozlov, S. (eds) ATM Kinase. Methods in Molecular Biology, vol 1599. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6955-5_7

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  • DOI: https://doi.org/10.1007/978-1-4939-6955-5_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6953-1

  • Online ISBN: 978-1-4939-6955-5

  • eBook Packages: Springer Protocols

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